U.S. patent application number 12/753567 was filed with the patent office on 2010-10-07 for compositions of cholinesterase inhibitors.
This patent application is currently assigned to Colucid Pharmaceuticals, Inc.. Invention is credited to Marc W. Andersen, Daniel Coughlin, Shoucheng Du, Alexander Kolchinski.
Application Number | 20100256229 12/753567 |
Document ID | / |
Family ID | 42826715 |
Filed Date | 2010-10-07 |
United States Patent
Application |
20100256229 |
Kind Code |
A1 |
Du; Shoucheng ; et
al. |
October 7, 2010 |
COMPOSITIONS OF CHOLINESTERASE INHIBITORS
Abstract
The present invention is directed to compositions, methods of
use, and processes for the synthesis related to
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate, and its pharmaceutically acceptable salt forms,
including the hydrogen fumarate salt. The present invention also
relates to a novel form polymorph of
3-((S)-1-(dimethylamino)ethyl)phenylmethyl-((R)-1-phenylpropan-2
yl)carbamate, characterized by a unique X-ray diffraction pattern
and Differential Scanning Calorimetry profile, as well a unique
crystalline structure.
Inventors: |
Du; Shoucheng; (Pittsburgh,
PA) ; Andersen; Marc W.; (Raleigh, NC) ;
Coughlin; Daniel; (Hackettstown, NJ) ; Kolchinski;
Alexander; (Winchester, MA) |
Correspondence
Address: |
MINTZ, LEVIN, COHN, FERRIS, GLOVSKY AND POPEO, P.C
ONE FINANCIAL CENTER
BOSTON
MA
02111
US
|
Assignee: |
Colucid Pharmaceuticals,
Inc.
Durham
NC
|
Family ID: |
42826715 |
Appl. No.: |
12/753567 |
Filed: |
April 2, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61166143 |
Apr 2, 2009 |
|
|
|
Current U.S.
Class: |
514/487 ;
560/32 |
Current CPC
Class: |
A61P 25/16 20180101;
A61P 25/28 20180101; A61P 9/10 20180101; C07C 57/15 20130101; A61P
25/34 20180101; A61P 25/00 20180101; C07C 271/44 20130101; A61P
27/06 20180101; A61P 1/00 20180101 |
Class at
Publication: |
514/487 ;
560/32 |
International
Class: |
A61K 31/27 20060101
A61K031/27; C07C 271/44 20060101 C07C271/44; A61P 27/06 20060101
A61P027/06; A61P 25/16 20060101 A61P025/16; A61P 25/28 20060101
A61P025/28; A61P 1/00 20060101 A61P001/00; A61P 9/10 20060101
A61P009/10; A61P 25/34 20060101 A61P025/34 |
Claims
1. A hydrogen fumarate salt of compound 3d: ##STR00026##
characterized by an x-ray diffraction pattern substantially similar
to that set forth in FIG. 16.
2. A pharmaceutical composition comprising the hydrogen fumarate
salt of compound 3d: ##STR00027## and a pharmaceutically acceptable
carrier or excipient.
3. The composition according to claim 2, wherein the compound has a
purity of greater than 98.0% as determined by LCMS.
4. The composition according to claim 2, wherein the compound
contains less than 2% impurity.
5. The composition according to claim 2, wherein the compound
contains less than 2% d-methamphetamine.
6. A method of treating or preventing a nervous system condition,
cholinergic deficiency or glaucoma in an individual by
administering a composition according to claim 2.
7. The method of claim 6, wherein said nervous system condition is
selected from a central nervous system condition, a peripheral
nervous system condition, and autonomic nervous system
condition.
8. The method of claim 6, wherein said central nervous system
condition is selected from Parkinson's disease, memory impairment,
and cognitive impairment.
9. The method of claim 8, wherein said memory impairment is
selected from Alzheimer's disease, age-associated memory loss,
impairment in memory consolidation, impairment in short term
memory, mild cognitive impairment, and multiple sclerosis.
10. A method of treating or preventing a condition associated with
acetylcholinesterase activity in an individual by administering a
composition according to claim 2, wherein the condition is selected
from delayed gastric emptying, attention deficit hyperactivity
disorder (ADHD), phobia, stroke, multiple sclerosis, sleep
disorder, psychiatric disorder, pain, anticholinergic drug
overdose, tobacco dependence, and spasticity.
11. A method of promoting wakefulness in an individual by
administering a composition according to claim 2.
12. The method of claim 11, wherein the individual suffers from a
disorder or condition selected from wakefulness disorders,
hypersomnia, sleep apnea, sleep disorders of central origin,
fatigue, excessive daytime sleepiness associated with narcolepsy,
fatigue and excessive sleepiness associated with a depressive
disorder or with antidepressant therapy.
13. A process for preparing 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate hydrogen fumarate
comprising the steps of: (1) reacting L-methamphetamine with
carbonyldiimidazole to form
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide;
(2) reacting
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
with (S)-rivalphenol to form 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate; and (3) contacting
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate with fumaric acid to yield
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate hydrogen fumarate.
14. A compound having the structure ##STR00028## or a
pharmaceutically acceptable salt thereof.
Description
RELATED APPLICATION
[0001] This application claims priority under 35 U.S.C. .sctn.119
to U.S. Ser. No. 61/166,143, filed Apr. 2, 2009, the contents of
which are incorporated herein.
FIELD OF THE INVENTION
[0002] The present invention is directed to compositions, methods
of use and processes for the synthesis of substantially pure
cholinesterase inhibitors, specifically
3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2-yl)carbamate, and its pharmaceutically
acceptable salt forms, including the hydrogen fumarate salt. The
invention is also directed to certain polymorphs and methods of
using such polymorphs.
BACKGROUND OF THE INVENTION
[0003] Cholinesterase inhibitors (ChEIs) of the carbamate type
("stigmines") are known. Towards the end of the nineteenth century,
physostigmine found medicinal use in the treatment of glaucoma.
More recently, physostigmine has been used for treatment of
myasthenia gravis and Alzheimer disease. However, the severity of
the side effects associated with high doses of physostigmine has
spurred the search for other carbamate cholinesterase inhibitors
that are safer and better tolerated. Examples of such stigmines are
the approved AD drug rivastigmine (marketed as Exelon.RTM.) and the
experimental AD drug phenserine. Although these second generation
cholinesterase inhibitors are better tolerated than physostigmine,
their application is still hampered by limited efficacy and a
narrow therapeutic window. Shown below are representative examples
of carbamate cholinesterases ("stigmines"):
##STR00001##
[0004] Stigmines inhibit AChE by transferring their carbamoyl group
to a serine residue in the active site (semi-irreversible
inhibition, see the schematic mechanism shown below). The
covalently bound carbamate is slowly hydrolyzed to reconstitute the
active enzyme. During this process, a carbamic acid is released
that in turn dissociates into carbon dioxide and an amine. In known
stigmines, this amine is a small molecule that is considered
pharmacologically inactive. We hypothesized that this mechanism
could be leveraged to release a biologically active amine during
the process of carbamic acid dissociation. Thus, these stigmines
could provide both cholinesterase inhibition and actions at
additional relevant targets in a single molecule, potentially
leading to increased efficacy and tolerability compared to known
cholinesterase inhibitors. Shown below is a schematic mechanism of
action for cholinesterase inhibition by carbamates:
##STR00002##
[0005] Recently, the concept of providing ChEIs with additional
pharmacology has received increased attention.
SUMMARY OF THE INVENTION
[0006] The present invention relates to a hydrogen fumarate salt of
compound 3d:
##STR00003##
characterized by an x-ray diffraction pattern substantially similar
to that set forth in FIG. 16.
[0007] The present invention relates to a pharmaceutical
composition comprising the hydrogen fumarate salt of compound
3d:
##STR00004##
and a pharmaceutically acceptable carrier or excipient.
[0008] In one aspect of the invention, the compound has a purity of
greater than 98.0% as determined by LCMS. In one aspect of the
invention, the compound contains less than 2% impurity. In one
aspect of the invention, the compound contains less than 2%
d-methamphetamine.
[0009] The present invention relates to a method of treating or
preventing a nervous system condition, cholinergic deficiency or
glaucoma in an individual by administering a composition of the
invention. The present invention relates to the use of a
composition of the invention in the manufacture of a medicament for
treating or preventing an individual having a nervous system
condition, cholinergic deficiency or glaucoma. In one aspect, the
nervous system condition is selected from a central nervous system
condition, a peripheral nervous system condition, and autonomic
nervous system condition. In one aspect, the central nervous system
condition is selected from Parkinson's disease, memory impairment,
and cognitive impairment.
[0010] In one aspect, the memory impairment is selected from
Alzheimer's disease, age associated memory loss, impairment in
memory consolidation, impairment in short term memory, mild
cognitive impairment, and multiple sclerosis.
[0011] The present invention relates to a method of treating or
preventing a condition associated with acetylcholinesterase
activity in an individual by administering a composition of the
invention, wherein the condition is selected from delayed gastric
emptying, attention deficit hyperactivity disorder (ADHD), phobia,
stroke, multiple sclerosis, sleep disorder, psychiatric disorder,
pain, anticholinergic drug overdose, tobacco dependence, and
spasticity. The present invention relates to the use of a
composition of the invention in the manufacture of a medicament far
treating or preventing an individual having a condition associated
with acetylcholinesterase activity, wherein the condition is
selected from delayed gastric emptying, attention deficit
hyperactivity disorder (ADHD), phobia, stroke, multiple sclerosis,
sleep disorder, psychiatric disorder, pain, anticholinergic drug
overdose, tobacco dependence, and spasticity.
[0012] The present invention relates to a method of promoting
wakefulness in an individual by administering a composition of the
invention. The present invention relates to the use of a
composition of the invention in the manufacture of a medicament for
promoting wakefulness in an individual. In one aspect, the
individual suffers from a disorder or condition selected from
wakefulness disorders, hypersomnia, sleep apnea, sleep disorders of
central origin, fatigue, excessive daytime sleepiness associated
with narcolepsy, fatigue and excessive sleepiness associated with a
depressive disorder or with antidepressant therapy.
[0013] The present invention relates to methods, wherein the
individual is a human.
[0014] The present invention relates to a process for preparing
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate hydrogen fumarate comprising the steps of: (1)
reacting L-methamphetamine with carbonyldiimidazole to form
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide;
(2) reacting
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
with (S)-rivalphenol to form 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate; and (3) contacting
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate with fumaric acid to yield
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate hydrogen fumarate.
[0015] The present invention relates to a compound having the
structure
##STR00005##
or a pharmaceutically acceptable salt thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] FIG. 1 is a table showing a comparison of three salt forms
of compound 3d.
[0017] FIG. 2 is two microscopic views showing hygroscopicity
testing of the hydrogen sulfate salt of compound 3d which
deliquesces after 3 days at 40.degree. C./75 RH. The views A-1 and
A-2 show the appearance after three and seven days at 75% RH,
respectively.
[0018] FIG. 3 is two microscopic views showing hygroscopicity
testing of the hydrogen fumarate salt of compound 3d which is a
solid after 7 days at 40.degree. C./75 RH. Analysis of compound 3d
hydrogen fumarate by microscopy indicates that the salt remains
crystalline with traces of amorphous material after 7 days at 75%
relative humidity.
[0019] FIG. 4 is photograph showing a gelatin capsule, size #000
filled with 200 mg of compound 3d hydrogen fumarate. The API fit in
to size #000 with minimal effort and manual packing. This indicates
that 200 mg of API can fit into the next smallest capsule size
#00.
[0020] FIG. 5 is a HPLC trace showing the results of chiral HPLC
separation for all possible isomers of compound 3d. No isomer
impurities are observed in batches of compound 3d.
[0021] FIG. 6 is an HPLC trace showing the results of forced
degradation of compound 3d in solution under an ultraviolet lamp
for 2 hours 45 min.
[0022] FIG. 7 is a table showing the results of a salt selection
study of 15 salts of compound 3d using IPA as a solvent.
[0023] FIG. 8 is a table summarizing melting point data for the
hydrogen succinate salt of compound 3d.
[0024] FIG. 9 is a table showing a comparison of solid state
properties of samples of hydrogen fumarate and hydrogen sulfate
samples of compound 3d.
[0025] FIG. 10 is a table showing a comparison of hygroscopicity of
the hydrogen fumarate and hydrogen sulfate salt forms of compound
3d.
[0026] FIG. 11 is a graph showing the sorption isotherm of compound
3d. Compound 3d does not absorb much water during absorption phase;
maximum moisture uptake is about 1% (at 75 RH). Adsorption appears
reversible.
[0027] FIG. 12 is a table showing the solid state stability of the
hydrogen fumarate salt of compound 3d. Samples of compound 3d
hydrogen fumarate were stable to heat (40.degree. C.) for 7 days
and to UV (220 W-hr/m.sup.2) and visible light for
7.3.times.10.sup.6 lux hours.
[0028] FIG. 13 is two microscopic views of the hydrogen fumarate
salt of compound 3d under mechanical stress. Micronizing a sample
of the fumarate salt of compound 3d for 20 minutes at 30 Hz
resulted in "gumming" after standing for 20 minutes.
[0029] FIG. 14 is a table showing the chemical stability of the
hydrogen fumarate salt of compound 3d at 50 min, 2 hour, 24 hour,
and 48 hour time points.
[0030] FIG. 15 is a table showing the chemical stability of the
hydrogen fumarate salt of compound 3d at 24 hour, 48 hour, 5 day
and 7 day time points. The hydrogen fumarate salt of compound 3d
was stable in water for up to 7 days at 40.degree. C.
[0031] FIG. 16 is an X-ray diffractogram for the hydrogen fumarate
salt of compound 3d.
[0032] FIG. 17 is a DSC thermogram for the hydrogen fumarate salt
of compound 3d.
[0033] FIG. 18 is a X-ray diffractogram for the hydrogen succinate
salt of compound 3d.
[0034] FIG. 19 is a DSC thermogram for the hydrogen succinate salt
of compound 3d.
[0035] FIG. 20 is a X-ray diffractogram for the hydrogen sulfate
salt of compound 3d.
[0036] FIG. 21 is a DSC thermogram for the hydrogen sulfate salt of
compound 3d.
[0037] FIG. 22 is a .sup.1HNMR spectrum of the free base of
compound 3d.
[0038] FIG. 23 is a .sup.1HNMR spectrum of the hydrogen fumarate
salt of compound 3d.
[0039] FIG. 24 is a .sup.1HNMR spectrum of the hydrogen sulfate
salt of compound 3d.
[0040] FIG. 25 is a .sup.1HNMR spectrum of the hydrogen succinate
salt of compound 3d.
[0041] FIG. 26 is an .sup.1HNMR spectrum of a urea intermediate
isolated during the preparation of compound 3d.
[0042] FIG. 27 is a bar graph that shows the step-through latencies
in a rat passive avoidance model). Rats received a dose of 0.2
mg/kg scopolamine s.c. 40 minutes before training and doses of 0,
8, 12, 16, or 24 mg of 4a i.p. 30 minutes before training (Example
6).
[0043] FIG. 28 is a graph that shows total minutes of sleep loss
versus dose of compounds of the invention, modafinil,
d-amphetamine, and rivastigmine (Example 12).
[0044] FIG. 29 is a graph that shows that rebound hypersomnolence
is not observed following administration of Compound B (Example
12).
[0045] FIG. 30 is a graph that shows the Compound B did not cause
an increase in body temperature (Example 12).
[0046] FIG. 31 is a graph that shows that Compound B did not cause
hyperactivity indicating an absence of stimulant activity (Example
12).
DETAILED DESCRIPTION OF THE INVENTION
[0047] The features and other details of the invention, either as
steps of the invention or as combinations of parts of the
invention, will now be more particularly described and pointed out
in the claims. It will be understood that the particular
embodiments of the invention are shown by way of illustration and
not as limitations of the invention. The principle features of this
invention can be employed in various embodiments without departing
from the scope of the invention.
[0048] The present invention is directed to a composition
comprising a pharmaceutically acceptable salt of compound 3d:
##STR00006##
In one aspect, the composition of the invention is the hydrogen
fumarate salt of compound 3d. In one aspect, the composition of the
invention is the hydrogen succinate salt of compound 3d. In one
aspect, the composition of the invention is the hydrogen sulfate
salt of compound 3d.
[0049] In one aspect, the composition of the invention comprises
compound 3d having a purity greater than 98.0% as determined by
LCMS. In one aspect, the composition of the invention comprises
compound 3d having a purity of greater than 99.0% as determined by
LCMS. In one aspect, the composition of the invention comprises
compound 3d having a purity of greater than 99.5% as determined by
LCMS. In one aspect, the composition of the invention comprises a
salt of compound 3d having a purity of greater than 99.7% as
determined by LCMS.
[0050] In one aspect, the composition of the invention comprises a
salt of compound 3d having a purity of greater than 98.0% as
determined by LCMS. In one aspect, the composition of the invention
comprises a salt of compound 3d having a purity of greater than
99.0% as determined by LCMS. In one aspect, the composition of the
invention comprises a salt of compound 3d having a purity of
greater than 99.5% as determined by LCMS. In one aspect, the
composition of the invention comprises a salt of compound 3d having
a purity of greater than 99.7% as determined by LCMS.
[0051] In one aspect, the composition of the invention contains
less than 2% impurity. In one aspect, the composition of the
invention contains less than 1% impurity. In one aspect, the
composition of the invention contains less than 0.5% impurity. In
one aspect, the composition of the invention contains less than
0.1% impurity. In one aspect, the composition of the invention
contains 0% impurity.
[0052] In one aspect, the composition contains less than 2%, less
than 1%, less than 0.5%, less than 0.1%, or 0% of an impurity such
as unreacted starting matter or a by-product formed during the
procedure to make the compound. Examples of impurities include
activated urea:
##STR00007##
rivaphenol, L-methamphetamine, D-methamphetamine, and symmetrical
urea
##STR00008##
[0053] The present invention is directed to a composition of the
invention further comprising a pharmaceutically acceptable carrier
or excipient.
[0054] The present invention is directed to certain polymorphs of
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate, including polymorphs of its salt forms characterized
by X-ray diffraction pattern, DSC thermogram and/or crystal
structure, processes of making these polymorphs, pharmaceutical
compositions comprising these polymorphs, and methods of treating
or preventing a nervous system condition, cholinergic deficiency,
and glaucoma; methods of increasing acetylcholine; methods of
treating or preventing a condition associated with
acetylcholinesterase activity; and methods of promoting
wakefulness.
[0055] The present invention is directed to a hydrogen fumarate
salt of compound 3d:
##STR00009##
characterized by an X-ray diffraction pattern substantially similar
to that set forth in FIG. 16. The hydrogen fumarate salt of
compound 3d is characterized by an X-ray diffraction pattern
including characteristic peaks at about 15.3, 17.0, 18.8, 21.3,
23.2, 23.6, 25.7, and 30.1 degrees 2-theta. The hydrogen fumarate
salt of compound 3d is further characterized by an X-ray
diffraction pattern including characteristic peaks at about 9.8,
10.6, 11.9, 13.8, 15.3, 17.0, 18.8, 21.3, 23.2, 23.6, 25.7, 26.4,
28.3, and 30.1 degrees two-theta.
[0056] The present invention is also directed to a hydrogen
fumarate salt of compound 3d characterized by a Differential
Scanning Calorimetry (DSC) thermogram having a single maximum value
at about 100.35, as measured by a DSC Q100 V9.8 Build 96
instrument.
[0057] A further embodiment of the present invention is a hydrogen
fumarate salt of compound 3d characterized by an X-ray diffraction
pattern substantially similar to that set forth in FIG. 16 and
further characterized by a Differential Scanning Calorimetry (DSC)
thermogram having a single maximum value at about 100.35, as
measured by a DSC Q100 V9.8 Build 96 instrument.
[0058] Another embodiment of the present invention is a hydrogen
fumarate salt of compound 3d characterized by an X-ray diffraction
pattern including characteristic peaks at about 15.3, 17.0, 18.8,
21.3, 23.2, 23.6, 25.7, and 30.1 degrees 2-theta and further
characterized by a Differential Scanning Calorimetry (DSC)
thermogram having a single maximum value at about 100.35, as
measured by a DSC Q100 V9.8 Build 96 instrument.
[0059] Yet another embodiment of the present invention is a
hydrogen fumarate salt of compound 3d characterized by an X-ray
diffraction pattern including characteristic peaks at about 9.8,
10.6, 11.9, 13.8, 15.3, 17.0, 18.8, 21.3, 23.2, 23.6, 25.7, 26.4,
28.3, and 30.1 degrees two-theta and further characterized by a
Differential Scanning Calorimetry (DSC) thermogram having a single
maximum value at about 100.35, as measured by a DSC Q100 V9.8 Build
96 instrument.
[0060] The present invention is directed to a hydrogen fumarate
salt of compound 3d produced by a process comprising the steps
of:
[0061] (1) reacting L-methamphetamine with carbonyldiimidazole to
form
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide;
[0062] (2) reacting
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
with (S)-rivalphenol to form 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate; and
[0063] (3) contacting 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate with fumaric acid to
yield 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate hydrogen fumarate. A
further embodiment of the invention is a hydrogen fumarate salt of
compound 3d produced by the aforementioned process and further
characterized by an X-ray diffraction pattern substantially similar
to that set forth in FIG. 16. Another embodiment of this invention
is a hydrogen fumarate salt of compound 3d produced by the
aforementioned process and further characterized by an X-ray
diffraction patter including characteristic peaks at about 15.3,
17.0, 18.8, 21.3, 23.2, 23.6, 25.7, and 30.1 degrees two-theta. Yet
another embodiment of the invention is a hydrogen fumarate salt
produced by the aforementioned process and further characterized by
an X-ray diffraction pattern including characteristic peaks at
about 9.8, 10.6, 11.9, 13.8, 15.3, 17.0, 18.8, 21.3, 23.2, 23.6,
25.7, 26.4, 28.3, and 30.1 degrees two-theta. Another embodiment of
the invention is a hydrogen fumarate salt of compound 3d produced
by the aforementioned process and further characterized by a DSC
thermogram having a single maximum value at about 100.35, as
measured by a DSC Q100 V9.8 Build 96 instrument.
[0064] A further embodiment of the invention is a hydrogen fumarate
salt produced by the aforementioned process, further characterized
by an X-ray diffraction pattern substantially similar to the set
forth in FIG. 16 and by a DSC thermogram having a single maximum
value at about 100.35, as measured by a DSC Q100 V9.8 Build 96
instrument.
[0065] Another embodiment of the invention is a hydrogen fumarate
salt produced by the aforementioned process, further characterized
by an X-ray diffraction pattern including characteristic peaks at
about 15.3, 17.0, 18.8, 21.3, 23.2, 23.6, 25.7, and 30.1 degrees
two-theta and by a DSC thermogram having a single maximum value at
about 100.35, as measured by a DSC Q100 V9.8 Build 96
instrument.
[0066] Yet another embodiment of the invention is a hydrogen
fumarate salt produced by the aforementioned process, further
characterized by an X-ray diffraction pattern including
characteristic peaks at about 9.8, 10.6, 11.9, 13.8, 15.3, 17.0,
18.8, 21.3, 23.2, 23.6, 25.7, 26.4, 28.3, and 30.1 degrees
two-theta and by a DSC thermogram having a single maximum value at
about 100.35, as measured by a DSC Q100 V9.8 Build 96
instrument.
[0067] The present invention is also directed at a hydrogen
fumarate salt of compound 3d produced by a process comprising the
step of recrystallizing a crude preparation of the hydrogen
fumarate salt of compound 3d from an organic solvent or a mixture
of an organic solvent and water. In particular embodiment, the
hydrogen fumarate salt of compound 3d is produced by a process
comprising the step of recrystallizing a crude preparation of the
compound 3d from an organic solvent. The organic solvent may be an
acetate such as isopropyl acetate (IPA). In another embodiment, the
hydrogen fumarate salt of compound 3d is produced by a process
comprising the step of recrystallizing a crude preparation of the
compound 3d from a mixture of isopropyl acetate and acetone.
[0068] The present invention is directed to a hydrogen sulfate salt
of compound 3d:
##STR00010##
characterized by an X-ray diffraction pattern substantially similar
to that set forth in FIG. 20. The present invention is directed to
a hydrogen sulfate salt of compound 3d characterized by an x-ray
diffraction pattern including characteristic peaks at about 10.1,
13.9, 14.9, 15.7, 18.1, 19.4, 19.9, 20.2, 21.3, 23.8, 24.2, and
26.1 degrees two-theta. The present invention is directed to a
hydrogen sulfate salt of compound 3d:
##STR00011##
characterized by an x-ray diffraction pattern including
characteristic peaks at about 10.1, 13.4, 13.9, 14.9, 15.7, 16.8,
17.1, 18.1, 19.4, 19.9, 20.2, 21.3, 22.2, 23.8, 24.2, 26.1, 26.6,
and 27.4 degrees two-theta.
[0069] The present invention is directed to a hydrogen sulfate salt
of compound 3d characterized by a Differential Scanning Calorimetry
(DSC) thermogram having a single maximum value at about 112.02, as
measured by a DSC Q100 V9.8 Build 96 instrument.
[0070] The present invention is directed to a hydrogen succinate
salt of compound 3d:
##STR00012##
characterized by an x-ray diffraction pattern substantially similar
to that set forth in FIG. 18. The present invention is directed to
a hydrogen succinate salt of compound 3d characterized by an x-ray
diffraction pattern including characteristic peaks at about 16.6.
16.9, 18.5, 19.1, 21.3, 23.4, 23.8, 25.6, and 29.0 degrees
two-theta.
[0071] The present invention is directed to a hydrogen succinate
salt of compound 3d characterized by an x-ray diffraction pattern
including characteristic peaks at about 10.4, 11.4, 14.9, 16.6.
16.9, 18.5, 19.1, 21.3, 23.4, 23.8, 25.6, 29.0, and 32.5 degrees
two-theta.
[0072] The present invention is directed to a hydrogen succinate
salt of compound 3d:
##STR00013##
[0073] characterized by a Differential Scanning Calorimetry (DSC)
thermogram having a single maximum value at about 79.07, as
measured by a DSC Q100 V9.8 Build 96 instrument.
[0074] The compositions of the present invention comprise a
compound or salt thereof that has cholinesterase activity. In one
aspect, the compound or salt inhibits a cholinesterase by competing
with a natural compound (e.g., acetylcholine (ACh) or
butyrylcholine (BuCh)) that binds cholinesterase. The
cholinesterase enzyme is inhibited when it is prevented from
inactivating a natural compound, such as the neurotransmitter ACh,
to any degree that cholinesterase would act on the neurotransmitter
in the absence of the compound. The cholinesterase inhibited can
be, for example, at least one member selected from the group
consisting of an acetylcholinesterase (AChe) or a
butyrylcholinesterase (BuChE). The compound or salt of the
invention can inhibit AChE alone or BuChE alone or can inhibit both
AChE and BuChE to similar or different degrees. AChE is located on
excitable membranes and inactivates ACh. The excitable membrane can
be a presynaptic neuron or a postsynaptic neuron. AChE is also
referred to as specific cholinesterase. BuChE is located on
excitable membranes and non-neuronal tissue such as blood cells.
BuChE is also referred to as pseudocholinesterase or nonspecific
cholinesterase. AChE and BuChE are regulators of cholinergic
neurotransmission in the central nervous system (brain and spinal
cord), peripheral nervous system and autonomic nervous system
(parasympathetic nervous system and sympathetic nervous
system).
[0075] A compound or salt of the invention can be useful as a CNS
active agent, or for cardiovascular therapy, or as an antibacterial
agent. For example, a CNS active agent is a useful for treating
Alzheimer's disease, Parkinson's disease, attention deficit
hyperactivity disorder (ADHD), depression, obsessive compulsive
disorders (OCD), anxiety disorders, chronic pain, or narcolepsy.
CNS active agents can be used to treat or prevent Alzheimer's
disease, neuropathic pain, spasticity, Parkinson's disease,
[0076] Administration of a composition of the invention can, for
example, result in an increase in ACh in the synapse of central
nervous system neurons which can compensate for the cholinergic
deficiency, for example, in Alzheimer's patients, thereby promoting
neuronal transmission to ultimately alleviate or ameliorate the
symptoms of Alzheimer's disease, Alzheimer's disease is accompanied
by symptoms that include cognitive impairment, disoriented
behavior, alter personality, difficulty speaking and comprehending
and impaired gait and movement. It has been suggested that
decreased cholinergic function is responsible for the symptoms of
Alzheimer's disease (Benzi, G., et al., European J. Pharmacol.
346:1-13 (1998); Korczyn, A. D., Exp. Opin. Invest. Drugs
9:2259-2267 (2000)).
[0077] A compound or salt of the invention can decrease the amount
of ACh synthesized or released, the inability of a neuron to
respond to ACh or inactivation of AChE. In Alzheimer's disease,
current treatments include the administration of compounds which
increase cholinergic signaling (Jann, M. W., Pharmacotherapy
20:1-12 (2000); Bachurin, S. O., Med. Res. Rev. 23:48-88 (2003)).
However, these compounds have modest efficacy, low response rate
(typically about 30%-50%) and numerous side effects such as nausea,
gastrointestinal problems and fatigue. In one embodiment, a
compound or salt of the invention inhibits AChE and increases
neurotransmitters, such as ACh, in the synapse of the central
nervous system neurons. Thus, for example, the compound or salt of
the invention inhibits AChE, which degrades ACh in the synapses of
neurons in Alzheimer's patients, and increase neurotransmitters in
the synapses.
[0078] Cholinergic deficiencies also characterize other disorders
such as Parkinson's disease, progressive supranuclear palsy,
vascular dementia and Down's syndrome (Korczyn, A. D., Exp. Opin.
Invest. Drugs 9:2259-2267 (2000)). Thus, the composition of the
invention can also be employed to increase the ACh in these
disorders.
[0079] Likewise, administration of a composition of the invention
can result in an increase in the neurotransmitter dopamine in the
central nervous system of patients with Parkinson's disease,
thereby promoting neuronal transmission to thereby diminish the
symptoms of Parkinson's disease.
[0080] In one embodiment, the compound or salt of the invention is
a memory-facilitating agent. In another embodiment, the compound or
salt of the invention is a cognition-facilitating agent.
[0081] The term "memory-facilitating agent," as used herein, refers
to a compound or salt that promotes memory in an individual,
prevents or minimizes a decline in memory in an individual or
participates in biological processes which are involved in memory
function.
[0082] The memory processes which can be facilitated by the
memory-facilitating agent can be memory consolidation, the process
of storing new information in long term memory ("Neuroscience:
Exploring The Brain," Bear, M. F. et al., Williams & Wilkins,
Baltimore, Md., Ch. 19, pp. 517-545 (1996); McGaugh, J. L. Science
287: 248-251 (2000), the teachings of which are hereby incorporated
by reference in their entirety); short-term memory (also referred
to as "working memory"), the process whereby newly acquired
information is maintained for short periods of time and the newly
acquired information is made available for further information
processing ("Neuroscience: Exploring The Brain," Bear, M. F. et
al., Williams & Wilkins, Baltimore, Md., Ch. 19, pp. 517-545
(1996); McGaugh, J. L. Science 287: 248-251 (2000); Becker, J. T.,
et al., Brain and Cognition 41:1-8 (1999), the teachings of which
are hereby incorporated by reference in their entirety);
declarative memory, which is the memory of facts and events
("Neuroscience: Exploring The Brain," Bear, M. F. et al., Williams
& Wilkins, Baltimore, Md., Ch. 19, pp. 517-545 (1996); McGaugh,
J. L. Science 287: 248-251 (2000); Tulving, E., et al., Science
247: 301-306 (1990); Squire, L. R., et al., Proc. Natl. Acad. Sci.
93: 13515-13522 (1996), the teachings of which are hereby
incorporated by reference in their entirety); procedural memory
(also referred to as "tacit knowledge" or "implicit knowledge"),
which is the memory for skills or behavior ("Neuroscience:
Exploring The Brain," Bear, M. F. et al., Williams & Wilkins,
Baltimore, Md., Ch. 19, pp. 517-545 (1996); McGaugh, J. L. Science
287: 248-251 (2000), the teachings of which are hereby incorporated
by reference in their entirety); or attention, acquisition,
retrieval or retention.
[0083] In another embodiment, the compound or salt is a
cognition-facilitating agent. The term "cognition-facilitating
agent," as used herein, refers to a compound or salt that promotes
activities associated with thinking, learning and acquiring
knowledge in an individual, prevents or minimizes a decline in
thinking, learning and acquiring knowledge in an individual or
participates in biological processes which are involved in
thinking, learning and acquiring knowledge. The decline in
thinking, learning and acquired knowledge (a cognitive disorder)
can be a consequence of or associated with another disease (e.g.,
Alzheimer's disease) or condition of the central, or peripheral or
autonomic nervous system. The cognitive process that can be
facilitated by the cognition-facilitating agent can be assessed by
behavioral criteria and behavioral assays which, in turn, can
further define where, in the learning, thinking, and acquiring
knowledge process, the cognition-facilitating agents are
acting.
[0084] An "agent," as used herein, refers to a compound that can
produce a physical, chemical or biological effect that can be
stimulatory (e.g., an activating agent) or inhibitory (e.g., a
blocking agent). Agents that are stimulatory can be agonists.
Agents that are inhibitory can be antagonists or inverse agonists.
Inverse agonists are compounds that down-regulate receptor
activated activity thereby acting in a manner that is the opposite
of an agonist to the receptor. Thus, exposure or administration of
an inverse agonist can result in a diminished response compared to
exposure or administration of an agonist.
[0085] A cholinergic agent can be, for example, a compound that
stimulates the action of ACh thereby mediating ACh-mediated cell
signaling between two cells (a cholinergic agonist). Stimulation
can be, for example, a result of facilitating binding of ACh to a
cell surface receptor, interference with degradation of ACh,
stimulation of release of ACh, stimulation of synthesis of ACh,
activation of second messengers (e.g., phospholipase C, inositol
1,4,5-triphosphate, protein kinase C, protein kinase A) that
mediate ACh cell signaling, alteration of ion (e.g., sodium,
potassium) channels in target cells. An agent can also inhibit or
prevent any one or more of these effects (e.g., a cholinergic
antagonist).
[0086] The compound or salt of the invention can inhibit
cholinesterase activity, which can be expressed as an IC50. The
term "IC50," as used herein, refers to the concentration of a
compound that inhibits an activity or effect by 50%, e.g., by
reducing the frequency of a condition, such as memory or cognitive
loss by 50%; by reducing binding of a competitor molecule to a
protein (e.g., a receptor) by 50%; or by reducing the level of an
activity (e.g., cholinesterase activity) by 50%.
[0087] As used herein, an "individual" is any mammal. A mammal can
be a rodent (such as a rat, mouse or guinea pig), domesticated
animal (such as a dog or cat), ruminant animal (such as a horse or
a cow) or a primate (such as a monkey or a human). In a preferred
embodiment, the individual is a human.
[0088] An individual suffering from a condition can be treated by a
composition of the invention. For example, the condition can
include at least one condition selected from the group consisting
of a central nervous system condition, a peripheral nervous system
condition and an autonomic nervous system condition.
[0089] In a particular embodiment, the individual treated with a
composition of the invention has a central nervous system
condition. A "central nervous system condition," as used herein,
refers to any illness or ailment that affects the brain or spinal
cord of the individual. Central nervous system conditions treated
with a composition of the invention, can, for example, be a
consequence of a genetic disease, environmental exposure to a
compound or secondary to a primary illness or disease. The central
nervous system condition can be characterized by or a consequence
of inadequate neurotransmitter release, synthesis, processing,
re-uptake or cell signaling. The central nervous system condition
can additionally, or alternatively, be characterized by or a
consequence of failed or inadequate neuronal transmission due to
disruptions in ion channels.
[0090] In a particular embodiment, a central nervous system
condition is treated with a composition of the invention. The
composition of the invention can be used to treat conditions such
as depression, anxiety or mental conditions. Compounds of the
invention can be used to treat conditions such as Parkinson's
disease, a memory impairment or a cognitive impairment.
[0091] The memory impairments can be in a human individual. Memory
impairments that can be treated by the compounds of the invention
include Alzheimer's disease, age-associated memory loss, an
impairment in memory consolidation, an impairment in short term
memory, mild cognitive impairment, an impairment in declarative
memory and/or impairments in memory associated with or a
consequence of multiple sclerosis and/or Parkinson's disease.
[0092] The memory impairment treated by the compositions of the
invention can be a consequence of exposure to a muscarinic
cholinergic receptor antagonist. In one embodiment, the muscarinic
cholinergic receptor antagonist is atropine. In another embodiment,
the muscarinic cholinergic receptor antagonist is scopolamine. In
yet another embodiment, the muscarinic cholinergic receptor
antagonist is homatropine.
[0093] A muscarinic cholinergic receptor antagonist includes any
substance which blocks, diminishes, attenuates, inhibits, hinders,
limits, decreases, reduces, restricts or interferes with the action
of ACh thereby disrupting ACh-mediated cell signaling between
presynaptic and postsynaptic neurons. The antagonist can, for
example, oppose the action of ACh by acting in a manner which
prevents ACh from binding to a muscarinic cholinergic receptor on a
postsynaptic neuron, from mediating post-synaptic events following
binding of ACh to a muscarinic cholinergic receptor, interfere with
ACh degradation by acetylcholinesterase in the synaptic cleft or
interfere with release of ACh from presynaptic neurons.
[0094] In still another embodiment, the compositions of the
invention can be used to treat a peripheral nervous system
condition in an individual. The peripheral nervous system condition
can, for example, be a disease or illness consequent to or
associated with neurons which supply innervation to a skeletal
muscle (e.g., Myasthenia Gravis). Conditions of the peripheral
nervous system can be, for example, an impairment in the release of
acetylcholine from neurons at the neuromuscular junction of
skeletal, smooth or cardiac muscle.
[0095] The compositions of the invention can be used to treat an
autonomic nervous system condition (sympathetic nervous system,
parasympathetic nervous system) in an individual. The autonomic
nervous system conditions can be conditions which affect smooth
muscle of viscera, glands (endocrine glands, exocrine glands),
blood vessels or cardiac muscle. Autonomic nervous system
conditions treated employing the compounds of the invention can be
post-operative distension and urinary retention. Conditions of the
autonomic nervous system can be an impairment in a function
associated with the autonomic nervous system, for example, an
impairment in the release of norepinephrine from sympathetic
neurons or ACh from parasympathetic neurons at a synapse with a
cell (e.g., epithelial, nervous, muscle, connective tissue) in an
organ, blood vessel or gland. One skilled in the art would be
capable of diagnosing an individual with a central nervous system
condition, peripheral nervous system condition and an autonomic
nervous system condition.
[0096] An "impairment in memory or cognition," as used herein,
refers to a diminished capacity in memory and/or cognitive
processes in the human. The cognitive and/or memory processes and
impairments in cognitive and/or memory processes can be assessed or
determined by established techniques. For example, memory can be
assessed before, concomitantly with or after treatment of the
individual with a composition of the invention one or more well
established tests known to one of skill in the art. Such tests
include the Passive Avoidance Testing (Principles of
Neuropsychopharmacology), Feldman R. S., et al., Sinauer Assoc.,
Inc., Sunderland, Mass. (1997), the teachings of all of which are
incorporated by reference in their entirety); Rey Auditory Verbal
Learning Test (RAVLT) (L' examen clinique en psychologie), Rey A.,
Paris: Presses Universitaires de France (1964); a Wechsler Memory
Scale; Wechsler Memory Scale-Revised (Wechsler, D., Wechsler Memory
Scale-Revised Manual, NY, N.Y., The Psychological Corp. (1987));
California Verbal Learning Test-Second Edition (Delis, D. C., et
al., The Californian Verbal Learning Test, Second Edition, Adult
Version, Manual, San Antonio, Tex.: The Psychological Corporation
(2000)); Cognitive Drug Research (CDR) Computerized Assessment
Battery-Wesnes; Buschke's Selective Reminder Test (Buschke, H., et
al., Neurology 24: 1019-1025 (1974)); Brief Visuospatial Memory
Test-Revised; and Test of Everyday Attention (Perry, R. J., et al.,
Neuropsychologia 38: 252-271 (2000)).
[0097] In a particular embodiment, the memory of the human before,
during or after administration of the composition of the invention
is assessed or determined by a word recall test such as RAVLT.
[0098] In another embodiment, the invention described herein
provides a method of treating a nervous system condition in an
individual. The method includes administering to the individual a
composition of the invention. The compound of the composition
inhibits a cholinesterase thereby treating the nervous system
condition of the individual.
[0099] In a particular embodiment, administration of the compound
of the invention treats a central nervous system condition in an
individual. In one aspect, the compound inhibits
acetylcholinesterase thereby treating the central nervous system
condition in the individual. The compound, upon hydrolysis, e.g.,
by reaction with the acetylcholinesterase, becomes at least one
component of a pharmacologically active agent that further treats
the central nervous system condition in the individual.
[0100] A further embodiment of the invention is a method of
increasing acetylcholine in an in vitro sample. The method includes
administering to the in vitro sample a compound. The compound
inhibits a cholinesterase, thereby increasing acetylcholine in the
in vitro sample.
[0101] The in vitro sample can be a cell-free sample or a sample
containing cells. The cells employed can be mammalian cells (e.g.,
CHO cells), insect cells or bacterial cells. The method can be
employed to assess the ability of the compound to inhibit
cholinesterase and the pharmacologically active agent to affect
biological, chemical or physical processes prior to use in an
individual. The method can be packaged in a kit as an assay for
screening the compounds of the invention for cholinesterase
activity and pharmacological activity of the agents the compound
becomes upon hydrolysis.
[0102] Another embodiment of the invention is a method of
increasing acetylcholine in a tissue. The method includes
administering to the tissue a composition of the invention. The
compound of the composition inhibits a cholinesterase, thereby
increasing acetylcholine in the tissue and, upon hydrolysis, e.g.,
by reaction with the cholinesterase, becomes at least a component
of a pharmacologically active agent that further increases
acetylcholine in the tissue.
[0103] The tissue can be a nervous tissue, a muscle tissue
(cardiac, skeletal, smooth muscle) or a collection of any one or
more of a tissue type selected from the group consisting of nervous
tissue, muscle tissue, epithelial tissue and connective tissue. The
tissue can be isolated (removed from the individual).
[0104] An additional embodiment of the invention is a method of
increasing acetylcholine in an individual. The method includes
administering to the individual a composition of the invention in
the individual. The composition inhibits a cholinesterase (e.g.,
AChE, BuChE), thereby increasing acetylcholine.
[0105] In one embodiment, the compound or salt of the invention
increases acetylcholine in the central nervous system of the
individual. In another embodiment, the compound or salt of the
invention increases acetylcholine in the peripheral nervous system
of the individual. In yet another embodiment, administration of a
composition of the invention increases acetylcholine in the
autonomic nervous system of the individual. Techniques to assess
the increase of ACh in an in vitro sample, in a tissue and in an
individual are well-known to one skilled in the art. (See, for
example, Day, J. C., et al. Methods 23:21-39 (2001), the teachings
of which are hereby incorporated by reference in its entirety).
[0106] The further increase in acetylcholine can be an increase
mediated in a manner similar to the increase mediated by the
compound of the invention (inhibition of AChE) or an increase in
ACh by, for example, increasing the release of ACh, increasing the
synthesis of ACh or otherwise preventing the inactivation of
ACh.
[0107] In a further embodiment, the invention is a method of
increasing transmission between two or more neurons. The method
includes exposing the neurons to a compound of the invention. The
compound inhibits a cholinesterase, thereby increasing transmission
between the two or more neurons.
[0108] The transmission can be increased between two or more
neurons in vitro or in vivo. Techniques to determine an increase in
transmission in vitro and in vivo are well known to one skilled in
the art. For example, changes in depolarization of the
post-synaptic neuron can be recorded by electrophysiological
methods.
[0109] An increase in transmission in an individual can minimize or
alleviate central or peripheral nervous system conditions, such as
memory and cognitive impairments. For example, an increase in
cholinergic transmission (e.g., post-synaptic) in a human
individual can minimize or alleviate the symptoms associated with
Alzheimer's disease. An increase in dopaminergic transmission
(e.g., post-synaptic) in a human individual can minimize or
alleviate the symptoms associated with Parkinson's disease. The
compound of the invention can facilitate penetration of the
compound through the blood brain barrier, thereby permitting
delivery of a pharmacologically active agent, in particular, into
the central nervous system.
[0110] Another embodiment of the invention is a method of treating
a cholinergic deficiency in an individual. The method includes
administering to the individual a composition of the invention. The
composition of the invention inhibits a cholinesterase thereby
treating the cholinergic deficiency in the individual.
[0111] The cholinergic deficiency can be a nervous system
deficiency. For example, the composition of the invention can be
used to treat a human individual having Alzheimer's disease.
Presynaptic neurons degenerate rapidly in Alzheimer's disease which
limits the efficacy of ChE inhibition as the disease progresses
(Cutler, N. R., et. al. CNS Drugs 3:467-481 (1995)). ChE continues
to be present in the synapses of neurons in an individual with
Alzheimer's disease, hydrolyzing what little ACh may be present in
the synapse. Thus, the compounds of the invention can become a
cholinergic agonist thereby ameliorating the cholinergic deficiency
by increasing ACh-mediated synaptic transmission in the central
nervous system of individuals suffering from Alzheimer's disease,
mild cognitive impairment, age associated memory impairment, age
associated memory loss, natural aging, vascular dementia, dementia
with Lewis bodies and/or Parkinson's disease.
[0112] In an additional embodiment, the invention is a method of
treating an impairment in memory in an individual. The method
includes administering to the individual a composition of the
invention. The compound inhibits a cholinesterase thereby treating
the impairment in memory in the individual.
[0113] The memory impairment can be a memory impairment selected
from the group consisting of an impairment in memory consolidation,
an impairment in long-term memory and an impairment in short-term
memory. One skilled in the art would be capable of identifying an
individual with a memory impairment and assessing the
impairment.
[0114] In a particular embodiment, a human individual has an
impairment in memory associated with a condition selected from the
group consisting of Alzheimer's disease, Parkinson's disease,
age-associated memory loss, mild cognitive impairment and multiple
sclerosis.
[0115] In another embodiment the human individual treated with the
compound of the invention has age-related cognitive decline.
[0116] A further embodiment of the invention is a method of
delivering a pharmacologically active agent to a tissue. The term
"pharmacologically active agent," as used herein, refers to a
compound that influences biological processes by altering the
activity, localization and/or expression of molecules (e.g.,
neurotransmitters, peptides, proteins) which are directly or
indirectly involved in the biological processes. The method
includes administering to the tissue a compound of the invention.
The compound of the invention inhibits a cholinesterase and, upon
hydrolysis, e.g., by reaction with the cholinesterase, becomes at
least a component of a pharmacologically active agent, thereby
delivering the pharmacologically active agent to the tissue.
[0117] The tissue can be an in vitro tissue sample or can be a
tissue in vivo (in an individual). The tissue can be muscle tissue,
nervous tissue or any combination of muscle, nervous, connective or
epithelial tissue. The compound of the invention can be employed to
deliver a pharmacologically active agent to a tissue that is
proximal or distal to a tissue having a cholinesterase that is
inhibited by the compound of the invention. For example, a compound
of the invention can be employed to deliver a pharmacologically
active agent, such as a cholinergic agent, to a muscle tissue. The
compound of the invention can bind a cholinesterase
(acetylcholinesterase, butyrylcholinesterase) thereby inhibiting
the activity of the cholinesterase and, upon hydrolysis (with, for
example, a cholinesterase), become a cholinergic agent. The
pharmacologically active agent can be delivered to a muscle cell
proximate to the site of binding of the compound of the invention
to the cholinesterase or to a muscle cell distal to the site of
binding. Similarly, the compound can bind to a cholinesterase in a
neuron of the nervous system and deliver a cholinergic agent
proximal or distal to the site of binding.
[0118] The compound of the invention can bind to a cholinesterase
and, upon hydrolysis, e.g., by reaction with the cholinesterase,
deliver methamphetamine to a neuron proximate or distal to the site
of binding of the compound of the invention. Thus, the compounds of
the invention provide a method of delivering a pharmacologically
active agent to the central nervous system. The pharmacologically
active agents can diffuse to varying regions of the brain and
mediate their effects.
[0119] The invention includes a method of treating a condition of
an individual by administering a composition of the invention,
wherein the condition is a nervous system condition selected from
delayed gastric emptying, attention deficit hyperactivity disorder
(ADHD), phobia, sleep disorder, stroke, psychiatric disorder, pain,
anticholinergic drug overdose, tobacco dependence, Parkinson's
disease, memory impairment, and cognitive impairment. The invention
includes use of a composition of the invention in the manufacture
of a medicament for treating a condition of an individual, wherein
the condition is a nervous system condition selected from delayed
gastric emptying, attention deficit hyperactivity disorder (ADHD),
phobia, sleep disorder, stroke, psychiatric disorder, pain,
anticholinergic drug overdose, tobacco dependence, Parkinson's
disease, memory impairment, and cognitive impairment.
[0120] The invention includes a method of treating a condition in
an individual by administering a composition of the invention,
wherein the condition is selected from glaucoma, oncologic
condition, delayed gastric emptying, attention deficit
hyperactivity disorder (ADHD), phobia, stroke, multiple sclerosis,
sleep disorder, psychiatric disorder, pain, anticholinergic drug
overdose, tobacco dependence, and spasticity. The invention
includes use of a composition of the invention in the manufacture
of a medicament for treating a condition in an individual, wherein
the condition is selected from glaucoma, delayed gastric emptying,
attention deficit hyperactivity disorder (ADHD), phobia, stroke,
multiple sclerosis, sleep disorder, psychiatric disorder, pain,
anticholinergic drug overdose, tobacco dependence, and
spasticity.
[0121] In one embodiment, the invention includes a method of
treating a condition of an individual by administering a
composition of the invention, wherein the condition is selected
from delayed gastric emptying, attention deficit hyperactivity
disorder (ADHD), phobia, sleep disorder, stroke, psychiatric
disorder, pain, anticholinergic drug overdose, and tobacco
dependence. The invention includes use of a composition of the
invention in the manufacture of a medicament for treating a
condition of an individual, wherein the condition is selected from
delayed gastric emptying, attention deficit hyperactivity disorder
(ADHD), phobia, sleep disorder, stroke, psychiatric disorder, pain,
anticholinergic drug overdose, and tobacco dependence.
[0122] In one embodiment, the invention includes a method of
treating a condition of an individual by administering a
composition of the invention, wherein the condition is
anticholinergic drug overdose. The invention includes use of a
compound of the invention in the manufacture of a medicament for
treating a condition of an individual, wherein the condition is
anticholinergic drug overdose.
[0123] In one embodiment, the invention includes the methods
discussed above, wherein the individual is a human.
[0124] Another aspect of the invention includes a method of
promoting wakefulness in an individual by administering a
composition of the invention, wherein the individual suffers from a
disorder or condition selected from wakefulness disorders,
hypersomnia, sleep apnea, sleep disorders of central origin,
fatigue, excessive daytime sleepiness associated with narcolepsy,
fatigue and excessive sleepiness associated with a depressive
disorder or with antidepressant therapy.
[0125] Another aspect of the invention includes a method of
promoting wakefulness, thereby treating the individual for a
disorder or condition selected from a wakefulness disorder,
hypersomnia, sleep apnea, sleep disorder of central origin,
fatigue, excessive daytime sleepiness associated with narcolepsy,
fatigue and excessive sleepiness associated with a major depressive
disorder or with antidepressant therapy.
[0126] In one aspect, the invention includes a method for the
treatment of a wakefulness disorder by administering to composition
of the invention as a wake promoting agent. In one aspect, the
invention includes a method for the treatment of sleep apnea by
administering to an individual a composition of the invention as a
wake promoting agent. In one aspect, the invention includes a
method for the treatment of a sleep disorder of central origin by
administering to an individual a composition of the invention as a
wake promoting agent. In one aspect, the invention includes a
method for the treatment of fatigue by administering to an
individual a composition of the invention as a wake promoting
agent. In one aspect, the invention includes a method for the
treatment of excessive daytime sleepiness associated with
narcolepsy by administering to an individual a composition of the
invention as a wake promoting agent. In one aspect, the invention
includes a method for the treatment fatigue and excessive
sleepiness associated with a major depressive disorder by
administering to an individual a composition of the invention as a
wake promoting agent. In one aspect, the invention includes a
method for the treatment fatigue and excessive sleepiness
associated with antidepressant therapy.
[0127] Fatigue and excessive sleepiness are among the symptoms of a
major depressive disorder, and can be adverse experiences
associated with antidepressant therapy and are often residual
symptoms inadequately treated with SSRI antidepressant therapy.
Antidepressant therapy includes but is not limited to therapy with
the following antidepressants: tricyclic antidepressants, selective
serotonin reuptake inhibitors (SSRIs), serotonin and noradrenaline
reuptake inhibitors, monoamine oxidase inhibitors and monoamine
oxidase type A. In another aspect, antidepressant is selected from
citalopram, fluoxetine, fluoxetine hydrochloride, paroxetine,
paroxetine hydrochloride, and clomipramine hydrochloride.
[0128] In one aspect, the invention relates to hypersomnia, a
condition that is characterized by reoccurring episodes of
excessive daytime sleepiness (EDS) or prolonged nighttime sleep.
Different from feeling tired due to lack of or interrupted sleep at
night, persons with hypersomnia are compelled to nap repeatedly
during the day, often at inappropriate times such as at work,
during a meal, or in conversation. These daytime naps usually
provide no relief from symptoms. Patients often have difficulty
waking from a long sleep, and may feel disoriented. Other symptoms
may include anxiety, increased irritation, decreased energy,
restlessness, slow thinking, slow speech, loss of appetite,
hallucinations, and memory difficulty. Some patients lose the
ability to function in family, social, occupational, or other
settings. In one aspect, the invention includes a method for the
treatment of hypersomnia, which comprises administering to an
individual a composition of the invention as a wake promoting
agent. In another aspect, the invention includes a method for the
treatment of hypersomnia, which comprises administering to an
individual a composition of the invention as an arousing agent.
[0129] In another aspect, the invention includes a method of
promoting wakefulness, wherein the wakefulness disorder or
condition is selected from circadian rhythm disorder and fatigue
associated with multiple sclerosis.
[0130] In one aspect, the invention includes a method of promoting
wakefulness by administering a composition of the invention,
wherein the circadian rhythm disorder is selected from shift work
sleep disorder, sleep apnea, desynchronizing disorder in blind
individuals, time zone change syndrome, shift work sleep disorder,
irregular sleep pattern, delayed sleep syndrome, and advanced sleep
syndrome. In another aspect, the invention includes a method of
promoting wakefulness, wherein the circadian rhythm disorder is
selected from shift work sleep disorder, sleep apnea, and
desynchronizing disorder in blind individuals.
[0131] In one aspect, the invention relates to sleep apnea. Sleep
apnea is a sleep disorder characterized by pauses in breathing
during sleep. Each episode, called an apnea, lasts long enough so
that one or more breaths are missed, and such episodes occur
repeatedly throughout sleep. The standard definition of any apneic
event includes a minimum 10 second interval between breaths, with
either a neurological arousal (a 3-second or greater shift in EEG
frequency, measured at C3, C4, O1, or O.sub.2), a blood oxygen
desaturation of 3-4% or greater, or both arousal and desaturation.
Sleep apnea is diagnosed with an overnight sleep test called a
polysomnogram.
[0132] Clinically significant levels of sleep apnea are defined as
five or more episodes per hour of any type of apnea (from the
polysomnogram). There are three distinct forms of sleep apnea:
central, obstructive, and complex (i.e., a combination of central
and obstructive) constituting 0.4%, 84% and 15% of cases
respectively. Breathing is interrupted by the lack of respiratory
effort in central sleep apnea; in obstructive sleep apnea,
breathing is interrupted by a physical block to airflow despite
respiratory effort. In complex (or "mixed") sleep apnea, there is a
transition from central to obstructive features during the events
themselves.
[0133] In one aspect, the invention includes a method for the
treatment of sleep disorders of central origin by administering to
an individual a composition of the invention. In another aspect,
the invention includes a method for the treatment of sleep
disorders of central origin by administering to an individual a
composition of the invention, wherein the number of apneas
occurring during sleep apnea syndromes is reduced. In one aspect,
treatment of sleep disorders of central origin by administering a
composition of the invention contributes to improving diurnal
somnolence and the quality of nocturnal sleep.
[0134] In one aspect, the invention includes a method of promoting
wakefulness in an individual, wherein individual is being treated
for sleep apnea with CPAP. "CPAP" or "continuous positive airway
pressure" is a mechanical device for the treatment for sleep apnea
and other sleep-related breathing disorders (including snoring).
Treatment with a CPAP device is typically administered via the nose
or mouth of the patient.
[0135] Under CPAP treatment, a subject wears a tight-fitting
plastic mask over the nose when sleeping. The mask is attached to a
compressor, which forces air into the nose creating a positive
pressure within the subject's airways. The principle of the method
is that pressurizing the airways provides a mechanical "splinting"
action, which prevents or lessens airway collapse and therefore,
obstructive sleep apnea. Although an effective therapeutic response
is observed in most subjects who undergo CPAP treatment, many
subjects cannot tolerate the apparatus or pressure and refuse
treatment. Moreover, recent covert monitoring studies demonstrated
that long-term compliance with CPAP treatment is very poor. It is
known that subjects remove their mask while sleeping.
[0136] In another aspect, the invention relates to fatigue
associated with multiple sclerosis (MS). Multiple sclerosis is one
of the most common disabling neurologic diseases of young adults in
the United States, where an estimated 400,000 persons have the
disease. Although MS can cause a variety of disabling neurological
impairments such as blindness, paralysis, incoordination, and bowel
or bladder dysfunction, a less apparent symptom that can also be
severely disabling is fatigue. As used herein "fatigue" includes
loss of power, or capacity to respond to stimulation. Effect
treatment of such fatigue includes alleviating tiredness, or
sleepiness associated with multiple sclerosis and also promoting
wakefulness in multiple sclerosis individuals. The mechanism of MS
fatigue is poorly understood. It has been attributed to nerve
conduction abnormalities within the central nervous system and
increased energy demands caused by neurologic disability. Several
characteristics of MS fatigue are interference with physical
functioning and activities of daily living, aggravation by heat,
and worsening at the end of the day. In aspect, the invention
includes a method of treatment for fatigue associate with multiple
sclerosis, comprising administering to an individual a composition
of the invention to improve or prevent symptoms of multiple
sclerosis fatigue in the individual. In another aspect, the
invention includes alleviating tiredness, or sleepiness associated
with multiple sclerosis and also promoting wakefulness in multiple
sclerosis individuals.
[0137] One aspect of the invention includes a method for enhancing
alertness or increasing regularity of sleep rhythms in an
individual by administering a composition of the invention.
[0138] In one aspect, the invention includes a method of promoting
wakefulness, wherein the compound or salt of the composition
administered has a reduced abuse potential. In one aspect of the
invention, no psychostimulant-like effects are observed in the
individual following administration of the composition of the
invention. A psychostimulant is a drug that causes a sense of
well-being, decreases fatigue and depression, and increases the
desire to eat. Psychostimulant drugs can also cause mood changes
and trouble with sleeping. In another aspect of the invention, the
composition of the invention administered has a dose-limiting side
effect. In one aspect, of the invention, the side effect is
nausea.
[0139] In another aspect of the invention, administration of the
composition of the invention does not cause rebound hypersomnolence
in the individual. The term "hypersomnolence" refers to an
excessive need for sleep, especially during the day. "Idiopathic
hypersomnolence" means a need for excessive daytime sleep without a
known cause. In another aspect of the invention, administration of
the composition of the invention does not cause psychostimulant
effects.
[0140] Another aspect of the invention includes the manufacture of
a medicament for promoting wakefulness in an individual that
suffers from a disorder or condition selected from wakefulness
disorders, hypersomnia, sleep apnea, sleep disorders of central
origin, fatigue, excessive daytime sleepiness associated with
narcolepsy, fatigue and excessive sleepiness associated with a
depressive disorder or with antidepressant therapy, wherein said
medicament comprises a composition described herein.
[0141] Another aspect of the invention includes the manufacture of
a medicament for promoting wakefulness in an individual and thereby
treating the individual a disorder or condition selected from
wakefulness disorders, hypersomnia, sleep apnea, sleep disorders of
central origin, fatigue, excessive daytime sleepiness associated
with narcolepsy, fatigue and excessive sleepiness associated with a
depressive disorder or with antidepressant therapy, wherein said
medicament comprises a composition described herein.
[0142] The present invention is directed to a process for preparing
compound 3d: 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate comprising the steps
of:
[0143] (1) reacting L-methamphetamine with carbonyldiimidazole to
form
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide;
and
[0144] (2) reacting
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
with (S)-rivalphenol to yield 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate.
[0145] In one embodiment, the invention is a process for preparing
the hydrogen fumarate of compound 3d:
3-((S)-1-(dimethylamino)ethyl)phenyl methyl-((R)-1-phenylpropan-2
yl)carbamate hydrogen fumarate comprising the steps of:
[0146] (1) reacting L-methamphetamine with carbonyldiimidazole to
form
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide;
[0147] (2) reacting
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
with (S)-rivalphenol to yield 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate; and
[0148] (3) contacting 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate with fumaric acid to
yield 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate hydrogen fumarate.
[0149] In another embodiment, the invention is a process for
preparing 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate hydrogen succinate
comprising the steps of:
[0150] (1) reacting L-methamphetamine with carbonyldiimidazole to
form
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide;
[0151] (2) reacting
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
with (S)-rivalphenol to form 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate; and
[0152] (3) contacting 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate with succinic acid to
yield 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate hydrogen succinate.
[0153] In yet another embodiment, the invention is a process for
preparing 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate hydrogen sulfate
comprising the steps of:
[0154] (1) reacting L-methamphetamine with carbonyldiimidazole to
form
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide;
[0155] (2) reacting
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
with (S)-rivalphenol to form 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate; and
[0156] (3) contacting 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate with sulfuric acid to
yield 3-((S)-1-(dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate hydrogen sulfate.
[0157] In one aspect, the process comprising the step of
recrystallizing a crude preparation of the salt (e.g., fumarate,
sulfate, succinate) of compound 3d from an organic solvent or a
mixture of an organic solvent and water. In a particular
embodiment, the salt of compound 3d is produced by a process
comprising the step of recrystallizing a crude preparation of the
compound 3d from an organic solvent. The organic solvent may be an
acetate such as isopropyl acetate. In another embodiment, the salt
of compound 3d is produced by a process comprising the step of
recrystallizing a crude preparation of the compound 3d from a
mixture of isopropyl acetate (IPA) and acetone.
[0158] Compounds of the invention relate to rivastigmine-based
conjugates of amphetamine isomers. Rivastigmine was chosen as
starting point for the compounds because it has proven clinical
utility in the management of Alzheimer's disease and because the
corresponding phenol is chemically well characterized, accessible
and stable. Amphetamines were selected due to their attractive and
well characterized pharmacological properties. Amphetamines are
neurotransmitter uptake inhibitors (Creese, I. and S. D. Iversen,
The pharmacological and anatomical substrates of the amphetamine
response in the rat. Brain Res., 1975. 83: p. 419-436) with
stimulant properties (Wise, R. A. and P. P. Rompre, Brain dopamine
and reward Ann. Rev. Psychol., 1989. 40: p. 191-225; Mason, S. T.,
The neurochemistry and pharmacology of extinction behavior.
Neurosci. Biobehav. Rev., 1983. 7: p. 325-347) which could be
beneficial in the treatment of (geriatric) depression and fatigue,
two symptoms that often accompany Alzheimer's and Parkinson's
disease. In addition, dextro-amphetamine is a well known
cognitive
[0159] The bifunctional cholinesterase inhibitors of the invention
can be prepared through combination of a pharmacologically active
amine and the phenol moiety of a known cholinesterase inhibitor in
a single molecule. The resulting hybrid molecules retain their
ability to inhibit cholinesterase, both in vitro and in vivo, and
as demonstrated, release pharmacologically active amines following
decarbamylation of the inhibited enzyme.
[0160] The high level of brain and plasma cholinesterase inhibition
in absence of severe side effects following oral doses of a
compound or salt of the invention is unprecedented and suggests
that the bifunctional cholinesterase inhibitors of the invention
may have a greater therapeutic window than currently known
cholinesterase inhibitors. Since it can be expected that peripheral
cholinergic effects are antagonized by the release of adrenergic
agents, the increased tolerability may be a reflection of in vivo
pharmacological effects of released d-amphetamine. One aspect of
the present invention is directed to a compound having the
structure
##STR00014##
or a pharmaceutically acceptable salt thereof.
DEFINITIONS
[0161] As used herein, the phrase "pharmaceutically acceptable"
refers to those compounds, materials, compositions, and/or dosage
forms which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of human beings and
animals without excessive toxicity, irritation, allergic response,
or other problem or complication, commensurate with a reasonable
benefit/risk ratio.
[0162] As used herein, "pharmaceutically acceptable salts" refer to
derivatives of the disclosed compounds wherein the compound is
modified by making acid or base salts thereof. Examples of
pharmaceutically acceptable salts include, but are not limited to,
mineral or organic acid salts of basic residues such as amines;
alkali or organic salts of acidic residues such as carboxylic
acids; and the like. The pharmaceutically acceptable salts include
the conventional non-toxic salts or the quaternary ammonium salts
of the parent compound formed, for example, from non-toxic
inorganic or organic acids. For example, such conventional
non-toxic salts include, but are not limited to, those derived from
inorganic and organic acids selected from 2-acetoxybenzoic,
2-hydroxyethane sulfonic, acetic, ascorbic, benzene sulfonic,
benzoic, bicarbonic, carbonic, citric, edetic, ethane disulfonic,
ethane sulfonic, fumaric, glucoheptonic, gluconic, glutamic,
glycolic, glycollyarsanilic, hexylresorcinic, hydrabamic,
hydrobromic, hydrochloric, hydroiodide, hydroxymaleic,
hydroxynaphthoic, isethionic, lactic, lactobionic, lauryl sulfonic,
maleic, malic, mandelic, methane sulfonic, napsylic, nitric,
oxalic, pamoic, pantothenic, phenylacetic, phosphoric,
polygalacturonic, propionic, salicylic, stearic, subacetic,
succinic, sulfamic, sulfanilic, sulfuric, tannic, tartaric, and
toluene sulfonic.
[0163] The pharmaceutically acceptable salts of the present
invention can be synthesized from the parent compound that contains
a basic or acidic moiety by conventional chemical methods.
Generally, such salts can be prepared by reacting the free acid or
base forms of these compounds with a stoichiometric amount of the
appropriate base or acid in water or in an organic solvent, or in a
mixture of the two; generally, non-aqueous media like ether, ethyl
acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists
of suitable salts are found in Remington's Pharmaceutical Sciences,
18th ed., Mack Publishing Company, Easton, Pa., USA, p. 1445
(1990).
[0164] The term "wake promotion" or "promoting wakefulness" as used
herein, refers to a marked increase in the duration of wakefulness
of an individual. In one aspect, there is no rebound
hypersomnolence in an individual to whom a composition of the
invention is administered. In one aspect, there is a reduction in
drowsiness i.e., there is an increased state of mental alertness,
or the prevention of further progression into a deeper state of
drowsiness that prefaced administration of a composition of the
invention. The term "drowsiness" is art-recognized, including
decreased states of mental alertness.
[0165] As used herein, "treating" or "treatment" includes any
effect e.g., lessening, reducing, modulating, or eliminating, that
results in the improvement of the condition, disease, disorder,
etc. "Treating" or "treatment" of a disease state means the
treatment of a disease-state in a mammal, particularly in a human,
and include: (a) inhibiting an existing disease-state, i.e.,
arresting its development or its clinical symptoms; and/or (c)
relieving the disease-state, i.e., causing regression of the
disease state.
[0166] As used herein, "preventing" means causing the clinical
symptoms of the disease state not to develop i.e., inhibiting the
onset of disease, in a subject that may be exposed to or
predisposed to the disease state, but does not yet experience or
display symptoms of the disease state.
[0167] The following Examples are illustrative and should not be
interpreted in any way so as to limit the scope of the
invention.
EXAMPLES
Example 1
Synthesis of 3-((S)-1-(Dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate Free Base (3d) and
Conjugate Acid Salts (VII)
##STR00015##
[0168] Example 1A
Preparation of 3-((S)-1-(Dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate Free Base (3d)
##STR00016##
[0170] In a 22 L 4-neck flask equipped with addition funnel,
mechanical stirrer, "distillation head" with reflux condenser and
nitrogen inlet and a thermocouple 638 g (3.86 mol, 1.05 eq.) of
(S)-rivaphenol IV and 1791 g (5.5 mol, 1.5 eq.) cesium carbonate
were suspended in 9.2 L toluene and subsequently heated to reflux
for 30 min. The first distillate (.about.120 mL) contained about 1
mL of water and was discarded. Subsequently, 893 g (3.67 mol, 1.0
eq.) activated urea III (Example 1) was dissolved in 3.3 L of
toluene (heated to 40.degree. C. to facilitate dissolution) and
slowly added to the reaction mixture at a rate that maintained
adequate reflux (.about.1 h). The addition funnel was rinsed with
an additional 100 mL of toluene and this solution was added as well
over 10 min. After completed addition the reaction mixture was
maintained at reflux overnight. During this time an additional 1 mL
of water was collected in the distillation still trap. The reaction
mixture was cooled to 90.degree. C. before 1.1 L of 1N sodium
hydroxide solution was added. After heating for 3 h to 90.degree.
C. The mixture was allowed to cool to rt and transferred into an
extraction funnel. The separated toluene layer was washed with 1.6
L of 1N sodium hydroxide before being extracted with aqueous 1N HCL
(5.4.times.1 L+1.times.2.7 L). The combined acidic extracts were
then basified with 10% sodium hydroxide solution to reach pH 12-13
and subsequently extracted with TBME (1.times.1 L+1.times.5 L). The
combined ether extracts were washed with 11 L 1N sodium hydroxide,
two times 1.6 L of water, and 1.1 L of saturated brine before being
dried over sodium sulfate. Current chromatographic purity is 99.1%
AUC. Solvent evaporation followed by high vacuum drying over
weekend afforded 1150 g of the title compound VI (96%).
Example 1B
(R)--N-Methyl-N-(1-phenylpropan-2-yl)-1H-imidazole-1-carboxamide
(III)
##STR00017##
[0172] In a 3 L 1-neck flask with mechanical stirring, nitrogen
inlet and thermocouple 193.6 g (1.04 mol, 1.0 eq.)
L-methamphetamine hydrochloride I was suspended in 1.5 L of
pyridine (99+%, 0.1% water maximum) at rt (16.degree. C.).
Subsequently, 186.0 g (1.15 mol, 1.1 eq) carbonyldiimidazole (CDI,
II) was added in 5 portions over a period of 15 minutes. Upon
completion of the addition the temperature had dropped to
12-13.degree. C. The mixture was heated to 90.degree. C. over the
period of 1 hour and then kept at this temperature. LCMS control
after 1.5 h indicated almost complete conversion with 1.1% starting
material remaining. Addition of 8.4 g (0.052 mol, 0.05 eq) CDI and
heating to 90.degree. C. for an additional 0.5 h led to complete
conversion. The mixture was cooled to 40.degree. C. and stripped of
most of its solvent. The resulting suspension was taken into 1.5 L
of ethyl acetate and subsequently extracted with three 800 mL
portions of water/sat. NaHCO.sub.3 (7/1) to maintain an extraction
pH of 7-8. The combined aqueous extracts were extracted with 500 mL
of ethyl acetate. LCMS analysis showed .about.10% of the desired
product in the second organic layer. Both ethyl acetate phases were
combined and dried over magnesium sulfate. The solids were filtered
off and washed with little additional ethyl acetate. Evaporation of
the solvent under reduced pressure yielded 260 g (>100%) of a
yellow solid that contained some pyridine. LCMS analysis confirmed
a purity of 99.7%. For purification the yellow solid was suspended
in 450 mL of TBME and mechanically stirred for 2 hours to yield a
fine white solid in a yellow liquid. Upon addition of 1.5 L of
heptane the suspension was further stirred for 16 h before being
filtered. Washing with 500 mL of heptane yielded 246 g (97%) of the
desired compound III as a fine and slightly yellow solid. LCMS
analysis showed a purity of 99.8%. NMR showed 2.3 w/w % of
imidazole as impurity.
Example 1C
Preparation of 3-((S)-1-(Dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate Hydrogen Fumarate VIIa
(HX=Fumaric Acid)
##STR00018##
[0174] Fumaric acid was dissolved in acetone (5 L), containing 1 kg
(1.05 eq) of VI (993 g neat). The resulting solution was diluted
with 5 L of isopropyl acetate and mechanically stirred under
nitrogen. Massive precipitation was observed overnight. The
resulting precipitate was transferred to the Buchner funnels and
pressed on the filter funnel to remove 7 L of the mother liquor.
The precipitate was washed with 5 L of iPrOAc-acetone 1:1 mixture,
pressed on the filter and dried at 30.degree. in vacuum (-25 psi,
50 Torr) with gentle nitrogen flow for four days. The final weight
of the title compound 1.2 kg (90% v. free base).
[0175] The following data was consistent with the structure shown:
.sup.1H- and .sup.13C-NMR were consistent with structure. DSC
melting (onset): 101.7.degree. C.; LC/MS analysis indicated an
[M+H]=341.1 which is consistent with the molecular weight of the
free base portion VI. Elemental Analysis, calculated for
C.sub.21H.sub.28N.sub.2O.sub.2.times.C.sub.4H.sub.4O.sub.4 (456.53
g/mol): C, 65.77; H, 7.07; N, 6.14. Found: C, 65.61; H, 7.30; N,
5.99. Karl Fischer analysis for moisture indicated 0.18% water.
Example 1D
General Procedure for Preparing Salts VII from Compound 3d
[0176] Compound VI and the acid (HX) were combined in 10 parts of
tetrahydrofuran (2 moles of HX for every 1 mole of VI) and
concentrated to dryness. The resulting oil was slurried in 10 parts
of isopropylacetate overnight affording a solid precipitate which
was isolated by filtration. In some cases, repeating the
isopropylacetate slurry treatment was necessary to achieve a
consistently melting solid.
Example 1E
Preparation of 3-((S)-1-(Dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate Hydrogen Succinate VIIb
(HX=Succinic Acid)
##STR00019##
[0178] Using the general procedure 2.9 g (78%) of the title
compound VIIb was prepared as a white solid.
[0179] The following data was consistent with the structure shown:
.sup.1H- and .sup.13C-NMR were consistent with structure. DSC
melting (onset): 79.0.degree. C.; LC/MS analysis indicated an
[M+H]=341.1 which is consistent with the molecular weight of the
free base portion VI. Elemental Analysis, calculated for
C.sub.21H.sub.28N.sub.2O.sub.2.times.C.sub.4H.sub.6O.sub.4 (458.55
g/mol): C, 65.48; H, 7.47; N, 6.11. Found: C, 65.38; H, 7.50; N,
6.08. Karl Fischer analysis for moisture indicated 0.20% water.
Example 1F
Preparation of 3-(S)-1-(Dimethylamino)ethyl)phenyl
methyl-((R)-1-phenylpropan-2 yl)carbamate Hydrogen Sulfate VIIc
(HX=Sulfuric Acid)
##STR00020##
[0181] Using the general procedure 7.7 g (87%) of the title
compound VIIc was prepared.
[0182] The following data was consistent with the structure shown:
.sup.1H- and .sup.13C-NMR were consistent with structure. DSC
melting (onset): 112.5.degree. C.; LC/MS analysis indicated an
[M+H]=341.1 which is consistent with the molecular weight of the
free base portion VI. Elemental Analysis, calculated for
C.sub.21H.sub.28N.sub.2O.sub.2.times.H.sub.2SO.sub.4 (438.54
g/mol): C, 57.51; H, 76.90; N, 6.39; S, 7.31. Found: C, 57.66; H,
7.04; N, 6.31; S, 7.23. Karl Fischer analysis for moisture
indicated 0.47% water.
Example 2
Synthesis of Cholinesterase Inhibitors
##STR00021##
[0184] Synthesis of compounds 4a, 4b, 4c, and 4d was accomplished
as shown above. Thus, (-)-3-hydroxyphenylethyldimethylamine 1 was
reacted with carbonyl diimidazole in dry ethyl acetate to form the
activated imidazolide. Addition of the amphetamine isomers (2a-d)
gave access to the target carbamates 3a-d in good yields (40-60%
after purification by column chromatography). Free base carbamates
3a-d were converted into the corresponding hydrochloride salts 4a-d
by addition of hydrogen chloride in diethyl ether followed by
removal of the solvents. The identity and purity of the resulting
white solids was firmly established by .sup.1H-NMR and HPLC
analysis.
Example 3
In Vitro Inhibition of AChE and BuChE
[0185] The ability of compounds 4a, 4b, 4c, and 4d to inhibit AChE
(human recombinant) and BuChE (human, purified from erythrocytes)
was determined. Cholinesterase activity was determined
spectrophotometrically by a modified Ellman procedure (Ellman, GL,
A new and rapid colorimetric determination of acetylcholinesterase
activity. Biochem Pharmacol. 1961, July; 7:88-95). The results of
these experiments are provided in the table below and demonstrate
that carbamates 4a, 4b, 4c, and 4d were all more potent inhibitors
of acetylcholinesterase than the parent structure
(rivastigmine).
[0186] Carbamates 4a, 4b, 4c, and 4d had less affinity for
butyrylcholinesterase than rivastigmine. Consequently, all four
inhibitors were more selective AChE inhibitors than rivastigmine.
The increase in AChE selectivity was especially pronounced for
compounds 4a (incorporating d-amphetamine, which was 225-fold more
selective for AChE than rivastigmine) and 4d (incorporating
l-methamphetamine, which was 1200-fold more selective than
rivastigmine).
TABLE-US-00001 Inhibitory concentrations (IC50's) of
cholinesterases 4a-d and the parent structure rivastigmine against
recombinant human acetylcholinesterase and purified human
butyrylcholinesterase (plasma). rhAChE hBuChE Compound Amine (nM)
(nM) Selectivity Rivastigmine N/A 2615 179 1:15 4a d-amphetamine
508 7322 14:1 4b l-amphetamine 404 416 1:1 4c d-methamphetamine 131
334 3:1 4d l-methamphetamine 302 24400 81:1 Values represent the
mean of two or three independent experiments, each performed in
triplicate. Standard deviations were typically within 10% of the
IC.sub.50 value.
Example 4
In Vivo Inhibition of Cholinesterase
[0187] The in vivo effectiveness of the stigmine compounds in vivo
inhibitors was evaluated. Initially, the Maximum Tolerated Dose
(MTD) was determined for each compound. The MTD was defined as the
dose at which clear, yet reversible and non-life threatening
cholinergic effects were observed. The data in the table below
shows that the compounds 4a, 4b, 4c, and 4d were better tolerated
than their parent structure rivastigmine.
TABLE-US-00002 Inhibition of total brain cholinesterase and total
plasma cholinesterase 60 minutes following oral dosing of compounds
4a-d and rivastigmine at the Maximum Tolerated Dose or highest dose
tested. Plasma Brain Compound Amine MTD (mg/kg) ChEI ChEI
Rivastigmine.sup.# N/A 5 mg/kg 39% 56% 4a d-amphetamine >100
mg/kg 59% 72% 4b l-amphetamine 32 mg/kg 26% 25% 4c
d-methamphetamine >64 mg/kg 50% 50% 4d l-methamphetamine 10
mg/kg 45% 64% Values represent the mean of n = 4 animals per group.
Standard deviations were typically within 10% of the determined
cholinesterase activity. .sup.#Rivastigmine data is the mean of
three measurements between 30 and 180 min following dosing.
[0188] To establish the in vivo cholinesterase inhibition
properties of the compounds, rats were dosed orally with saline
solutions of the test compounds at the MTD (or at the highest dose
tested in case of 4a and 4e). Following administration, rats were
sacrificed and blood and brain samples were collected, processed
and ChE activity was determined spectrophotometrically as described
above. Initially, cholinesterase inhibition was quantified at 30,
60 and 180 min following oral dosing of rivastigmine to assess
time-dependence. As there was no consistent difference in the level
of inhibition achieved between these times (Bonferroni t-test),
which is in accord with reports of its long duration of
pseudo-irreversible inhibition, plasma and brain levels of
cholinesterase inhibition of all other compounds were determined at
a single time point (60 min). All compounds resulted in significant
inhibition of cholinesterase following oral dosing in both plasma
and brain, demonstrating the oral bioavailability and blood-brain
barrier penetration of compounds 4a-d.
Example 5
Amphetamine Release
[0189] Next, the release of amphetamine from inhibited
cholinesterase was investigated indirectly by monitoring
reconstitution of enzyme activity as described above. Recombinant
human cholinesterase was incubated with an excess of carbamates
4a-d, resulting in >80% inhibition of enzyme activity. The
enzyme was separated from small organic molecules (carbamate and
possible degradation products) by size exclusion chromatography
over a Sephadex column. Purified, inhibited enzyme was incubated in
phosphate buffer at 37.degree. C., aliquots were drawn at various
time points and the enzymatic activity was determined as described
above. The data were plotted as the natural logarithm of the
percent inhibition versus time according to a first-order kinetics
model. The decarbamylation rate constant k was determined as the
slope of this line. The halflife t.sub.1/2 is directly related to k
(k=-ln(0.500)/t.sub.1/2).
[0190] The results of the decarbamylation experiments are presented
in the table below.
TABLE-US-00003 Decarbamylation rates (k) and decarbamylation
half-lives (t.sub.1/2) following inhibition of rhAChE by compounds
4a-d and rivastigmine k (h.sup.-1) t.sub.1/2 Compound Amine (h)
Rivastigmine N/A 0.007 >24 4a d-amphetamine 0.558 1.2 4b
l-amphetamine 0.057 12 4c d-methamphetamine 0.003 >24 4d
l-methamphetamine 0.011 >24
[0191] In order to unequivocally demonstrate that enzyme
reconstitution results in release of amphetamine, aliquots from the
decarbamylation experiments with 4a were analyzed for levels of
amphetamine using an LC/MS/MS method. It was demonstrated that
aliquots drawn at t=0 did not contain amphetamine, whereas aliquots
drawn 4 h later (corresponding to 80% reconstitution) contained 53
ng/mL of d-amphetamine. Similarly, plasma collected from rats one
hour following oral doses (64 mg/kg) of 4a contained significant
amounts of amphetamine (17 ng/mL).
Example 6
Memory Enhancement
[0192] To determine whether compound 4a had any memory enhancing
properties, its effects were tested in a scopolamine model of
passive avoidance (C. Bejar, R.-H. Wang, M. Weinstock, "Effects of
rivastigmine on scopolamine-induced memory impairment in rats,",
Eur. J. Pharmacol. 383, 1999, 231-240). Prior to training, rats
were injected with scopolamine hydrobromide (0.2 mg/kg, s.c, 40
minutes before training) and with either saline or compound 4a (8,
12, 16 or 24 mg/kg, i.p, 30 minutes prior to training). The animals
received a retention test 24 hours following training. The
retention test was identical to training except that no foot-shock
or drug was delivered. Latency to enter the dark chamber was
recorded.
[0193] Results (FIG. 27) demonstrated that pre-training
administration of scopolamine produced a robust and statistically
significant amnesia (p<0.0001). This amnesia could be alleviated
by doses of 8 and 12 mg/kg of compound 4a (p<0.01). These
results clearly demonstrate that compound 4a is able to improve
mnemonic performance in rats.
Example 7
Formulation Summary
[0194] Formulation studies indicate the compound 3d has favorable
properties for development as an oral drug. Solid compound 3d is
stable to heat (40.degree. C.) for 7 days, to UV for 220
W-h/m.sup.2, and no change after visible light exposure of
7.3.times.10 h equivalent. Compound 3d solutions are stable at pH
4, 5, 6, after 7 days based on HPLC recovery experiments. Compound
3d is highly soluble (>50 mg/mL) at pH 3-6. Stability indicating
HPLC method qualified for use in formulation work (linearity,
precision, accuracy, recovery).
[0195] Polymorph studies indicate that compound 3d is highly
crystalline and one form is predominant. Hygroscopicity testing at
40.degree. C./75 RH indicated stability over 7 days by differential
scanning calorimetry (DSC) and X-ray powder diffraction (XRPD)
studies. Mechanical stress ball pine indicates instability to
extreme thermal conditions (compound "gums"), likely due to
compound 3d melting point which is ca. 110.degree. C. Sixteen
vehicles for oral dosing (rodents) were tested and all behave
equally well (>50 mg/mL). Water, HMC, or HPMC (0.25, 0.5%) are
the preferred choices.
Example 8
Salt Selection
[0196] Batches of compound 3d were prepared as the free base.
Because the synthesis of compound 3d free base resulted in an oil,
which is not practical for development, focus shifted to
identifying a suitable a salt form. Fifteen salts of compound 3d
were initially screened. The initial salt selection screen resulted
in oils and gums. A second salt selection screen was conducted over
a 3 month period to screen 40 acids. The second salt selection
screen resulted in only gums/oils. Crystallization studies were
then conducted using "best solvents" not yet tried. The initial
crystallization study was repeated using the newly selected
solvents and avoiding heat due to the low melting point of compound
3d salts. Several solvents/conditions produced solids; isopropyl
acetate (IPA) was selected for further study. The top 10 preferred
salt forms were re-screened using IPA solvent. Three solid forms of
compound 3d were selected for further study. The 3 solid forms
chosen were succinate, fumarate, and sulfate salt (1:1 ratio). The
3 solid forms were screened for "druggable" properties: melting
point, crystallity, solubility, hygroscopicity, stability, etc. The
hydrogen succinate salt of compound 3d afforded a low melting point
(60-70.degree. C.) and amorphous character was shown by microscopy
and XRPD.
[0197] Further studies were conducted on the hydrogen fumarate (mp
110.degree. C.) and hydrogen sulfate (mp 120.degree. C.) salts of
compound 3d were conducted.
Example 9
Peak List for X-Ray Diffractogram of Hydrogen Fumarate Salt Shown
in FIG. 16
TABLE-US-00004 [0198] 2-Theta d(.ANG.) BG Height H % Area A % FWHM
4.221 20.9171 377 173 6.7 3363 5.1 0.165 8.451 10.4544 290 107 4.1
2422 3.7 0.193 9.510 9.2920 316 96 3.7 5616 8.5 0.498 9.759 9.0555
329 148 5.7 5616 8.5 0.322 10.670 8.2846 336 225 8.7 8415 12.8
0.317 11.919 7.4192 315 162 6.3 7377 11.2 0.387 13.891 6.3701 340
236 9.2 5509 8.4 0.198 15.319 5.7792 332 418 16.2 16246 24.7 0.331
16.641 5.3229 396 311 12.0 13291 20.2 0.364 17.010 5.2084 367 1712
66.4 63377 96.2 0.315 17.648 5.0215 478 89 3.5 1412 2.1 0.135
18.401 4.8176 548 218 8.5 5489 8.3 0.214 18.800 4.7163 533 1849
71.6 62424 94.8 0.287 19.431 4.5646 703 217 8.4 2718 4.1 0.107
20.412 4.3473 615 204 7.9 9152 13.9 0.381 20.641 4.2996 591 184 7.1
9152 13.9 0.424 21.340 4.1603 515 2581 100.0 65870 100.0 0.217
23.280 3.8178 438 541 21.0 22089 33.5 0.347 23.600 3.7668 456 635
24.6 24683 37.5 0.330 24.618 3.6133 460 88 3.4 3950 6.0 0.382
25.001 3.5588 424 119 4.6 3950 6.0 0.281 25.720 3.4610 425 1711
66.3 40559 61.6 0.201 26.421 3.3707 421 313 12.1 11227 17.0 0.305
27.801 3.2065 389 121 4.7 3260 4.9 0.229 28.340 3.1467 379 181 7.0
7421 11.3 0.348 28.681 3.1100 370 96 3.7 6372 9.7 0.563 30.129
2.9638 351 472 18.3 14090 21.4 0.254 30.729 2.9072 369 120 4.6 5042
7.7 0.358 31.550 2.8334 378 177 6.9 4838 7.3 0.233 32.148 2.7820
360 71 2.8 3089 4.7 0.370 32.500 2.7528 353 61 2.4 1500 2.3 0.208
34.156 2.6230 295 65 2.5 3724 5.7 0.487 34.480 2.5990 292 75 2.9
4629 7.0 0.522 36.500 2.4597 294 65 2.5 2600 3.9 0.338 37.809
2.3775 265 55 2.1 762 1.2 0.119 40.755 2.2122 273 62 2.4 847 1.3
0.117 41.930 2.1529 287 92 3.6 2322 3.5 0.214 43.560 2.0760 270 289
11.2 8615 13.1 0.253 45.590 1.9882 262 65 2.5 2741 4.2 0.356 48.057
1.8917 245 55 2.1 2607 4.0 0.400
Example 10
Peak List for X-Ray Diffractogram of Hydrogen Succinate Salt Shown
in FIG. 18
TABLE-US-00005 [0199] 2-Theta d(.ANG.) BG Height H % Area A % FWHM
4.190 21.0716 382 176 6.3 3738 3.5 0.181 8.430 10.4798 295 159 5.7
2712 2.6 0.145 10.360 8.5319 315 660 23.6 17735 16.8 0.229 11.410
7.7491 309 728 26.1 21540 20.4 0.251 13.600 6.5055 301 406 14.6
10266 9.7 0.215 14.071 6.2890 311 265 9.5 6278 5.9 0.202 14.900
5.9409 314 677 24.3 17805 16.9 0.223 15.860 5.5832 341 312 11.2
7850 7.4 0.214 16.620 5.3297 410 1072 38.5 42931 40.7 0.340 16.940
5.2296 389 1467 52.6 59059 56.0 0.342 17.650 5.0209 437 423 15.2
10329 9.8 0.208 18.550 4.7794 549 1023 36.7 21179 20.1 0.176 19.130
4.6357 437 2614 93.7 76323 72.3 0.248 19.470 4.5556 538 365 13.1
11293 10.7 0.263 20.149 4.4034 427 272 9.7 5527 5.2 0.173 20.830
4.2610 449 578 20.7 37689 35.7 0.554 21.260 4.1758 431 2789 100.0
105518 100.0 0.322 22.299 3.9834 429 235 8.4 5616 5.3 0.203 22.921
3.8769 489 265 9.5 5024 4.8 0.161 23.450 3.7906 421 670 24.0 58099
55.1 0.737 23.770 3.7403 450 1564 56.1 65703 62.3 0.357 24.280
3.6628 497 247 8.8 3483 3.3 0.120 25.560 3.4822 373 1635 58.6 68084
64.5 0.354 26.720 3.3336 354 344 12.3 11191 10.6 0.277 27.171
3.2793 340 263 9.4 14768 14.0 0.478 29.010 3.0755 321 934 33.5
33075 31.3 0.301 29.950 2.9810 348 256 9.2 6077 5.8 0.202 30.751
2.9052 353 94 3.4 3799 3.6 0.343 31.554 2.8331 371 106 3.8 1039 1.0
0.084 31.910 2.8023 322 294 10.5 10740 10.2 0.311 32.530 2.7503 353
527 18.9 15029 14.2 0.242 33.420 2.6790 327 131 4.7 5632 5.3 0.365
34.440 2.6020 316 108 3.9 3306 3.1 0.261 34.841 2.5729 300 114 4.1
3254 3.1 0.243 35.990 2.4934 282 58 2.1 1203 1.1 0.176 36.651
2.4499 266 105 3.8 6259 5.9 0.504 37.429 2.4008 267 116 4.2 3220
3.1 0.235 38.287 2.3489 267 75 2.7 2749 2.6 0.313 38.750 2.3219 257
57 2.0 3288 3.1 0.490 39.941 2.2554 256 212 7.6 10513 10.0 0.421
41.311 2.1837 279 176 6.3 6075 5.8 0.293 42.081 2.1455 297 99 3.5
1798 1.7 0.155 42.754 2.1133 304 58 2.1 958 0.9 0.141 43.359 2.0852
296 239 8.6 13269 12.6 0.471 44.501 2.0343 270 64 2.3 1649 1.6
0.220 46.019 1.9706 264 123 4.4 5328 5.0 0.369 46.812 1.9391 268
181 6.5 5336 5.1 0.251 47.833 1.9001 266 117 4.2 3651 3.5 0.266
49.130 1.8529 255 120 4.3 4282 4.1 0.302
Example 11
Peak List for X-Ray Diffractogram of Hydrogen Sulfate Salt Shown in
FIG. 20
TABLE-US-00006 [0200] 2-Theta d(.ANG.) BG Height H % Area A % FWHM
3.340 26.4331 533 3038 100.0 57856 90.0 0.162 6.667 13.2476 323 79
2.6 3559 5.5 0.384 10.061 8.7848 302 1035 34.0 20333 31.6 0.167
10.601 8.3388 291 342 11.3 7390 11.5 0.183 13.449 6.5782 340 611
20.1 12589 19.6 0.175 13.949 6.3435 382 1409 46.4 28394 44.2 0.171
14.370 6.1588 398 347 11.4 4731 7.4 0.116 14.921 5.9325 398 983
32.4 21041 32.7 0.182 15.340 5.7715 355 392 12.9 17597 27.4 0.382
15.770 5.6149 369 688 22.6 13593 21.1 0.168 16.391 5.4036 317 220
7.2 4587 7.1 0.177 16.869 5.2515 328 419 13.8 10637 16.5 0.216
17.160 5.1632 335 517 17.0 10469 16.3 0.172 17.540 5.0521 336 190
6.2 3438 5.3 0.154 18.060 4.9079 329 1825 60.1 38019 59.1 0.177
19.070 4.6502 423 459 15.1 13138 20.4 0.243 19.420 4.5671 465 2241
73.7 45484 70.8 0.173 19.930 4.4513 467 690 22.7 64278 100.0 0.792
20.230 4.3861 420 1698 55.9 62843 97.8 0.315 20.581 4.3121 428 149
4.9 1938 3.0 0.110 21.340 4.1603 329 1186 39.0 24848 38.7 0.178
22.210 3.9993 324 496 16.3 12758 19.8 0.219 22.700 3.9141 313 414
13.6 13033 20.3 0.267 23.851 3.7278 338 686 22.6 24796 38.6 0.307
24.150 3.6822 311 1003 33.0 52384 81.5 0.444 24.780 3.5901 348 372
12.3 7261 11.3 0.166 25.261 3.5228 341 258 8.5 4499 7.0 0.148
25.910 3.4359 321 500 16.5 27955 43.5 0.475 26.180 3.4011 341 693
22.8 20309 31.6 0.249 26.680 3.3385 329 458 15.1 8779 13.7 0.163
27.108 3.2868 375 142 4.7 1459 2.3 0.087 27.449 3.2467 314 452 14.9
16979 26.4 0.319 27.941 3.1907 303 313 10.3 17275 26.9 0.470 29.000
3.0765 309 187 6.2 3257 5.1 0.148 29.710 3.0046 304 226 7.4 8056
12.5 0.304 30.310 2.9465 286 326 10.7 13522 21.0 0.352 31.329
2.8529 256 79 2.6 2731 4.2 0.293 31.929 2.8006 250 282 9.3 5909 9.2
0.178 32.339 2.7661 253 68 2.2 1413 2.2 0.177 33.229 2.6940 269 155
5.1 5331 8.3 0.292 33.719 2.6560 258 127 4.2 7429 11.6 0.498 34.028
2.6325 274 100 3.3 3828 6.0 0.326 34.690 2.5838 257 310 10.2 8925
13.9 0.245 35.131 2.5524 254 159 5.2 3510 5.5 0.188 35.690 2.5137
259 222 7.3 6324 9.8 0.242 36.589 2.4539 270 107 3.5 3955 6.2 0.315
37.141 2.4188 277 58 1.9 3122 4.9 0.461 37.699 2.3842 255 182 6.0
5853 9.1 0.273 38.180 2.3553 244 106 3.5 3012 4.7 0.243 39.009
2.3071 257 81 2.7 1363 2.1 0.143 39.591 2.2745 244 90 3.0 3399 5.3
0.320 39.959 2.2544 259 162 5.3 4004 6.2 0.210 40.659 2.2172 245 84
2.8 1384 2.2 0.140 41.843 2.1572 252 69 2.3 2798 4.4 0.346 42.409
2.1297 259 129 4.2 4085 6.4 0.269 42.698 2.1159 263 80 2.6 2704 4.2
0.286 43.489 2.0792 259 174 5.7 6524 10.1 0.318 44.071 2.0531 253
125 4.1 3242 5.0 0.220 45.300 2.0002 260 66 2.2 1594 2.5 0.206
45.681 1.9844 244 75 2.5 6748 10.5 0.767 46.089 1.9678 250 66 2.2
2966 4.6 0.379 47.149 1.9260 234 89 2.9 3751 5.8 0.357 48.079
1.8909 243 67 2.2 757 1.2 0.095 48.941 1.8596 258 101 3.3 2501 3.9
0.210
Example 12
Wake Promotion
[0201] Certain clinical conditions are characterized by
unpredictable bouts of sleepiness that can interfere with the
ability to conduct activities of daily living, such as driving.
Examples are narcolepsy, and disturbances of diurnal rhythm, such
as adjustment to shift work. Currently approved therapies for such
conditions are amphetamines and modifenil. Significant limitations
of available therapies include rebound hypersomnolence and abuse
potential.
[0202] Test compounds in various dose ranges or vehicle were
administered to male Wistar rats 5 hours after lights on (CT-5).
EEG, EMG, locomotor activity, drink- and food-related activity, and
body temperature were concurrently monitored for 30 hr before and
after treatment from rats living in separate isolated recording
chambers. Sleep-wake discriminations were carried out using
SCORE2004.TM., proprietary real-time hardware and software
technology of Hypnion, Inc. Comparisons were made between the
reference compounds: d-amphetamine, rivastigmine, modafinil, and
the following test compounds:
TABLE-US-00007 Compound # Compound Name/Letter Code ##STR00022##
s-riva-l-amphetamine/A ##STR00023## s-riva-l-methamphetamine/B
##STR00024## s-riva-d-amphetamine/C ##STR00025##
s-riva-d-methamphetamine/D
[0203] Administration of d-amphetamine or modafinil increased the
duration of wakefulness (i.e., increased the total number of
minutes of sleep loss) in a dose-dependent manner. Although not
approved as a wake promoting agent, rivastigmine also increased
wakefulness. Higher doses of reference compounds were not tested
because of tolerability. Similarly, Compounds A, B, C and D caused
a dose-related increase in wakefulness. Of these, Compound B caused
an unexpectedly long increase in wakefulness that surpassed that
seen with the reference compounds tested (See, FIG. 28).
[0204] Unlike rivastigmine, rebound hypersomnolence was not
observed following administration of Compound B (See, FIG. 29).
This is an unexpected finding that would not be predicted from the
known actions of the compound's component stigmine or amine.
[0205] Compound B, also differed in other unexpected ways from the
reference compounds with respect to its effects on body temperature
and locomotor activity. Unlike d-amphetamine, Compound B did not
cause an increase in body temperature (hyperthermia), but rather
caused an opposite reduction in body temperature (hypothermia; See,
FIG. 30). Moreover, unlike d-methamphetamine, Compound B did not
cause locomotor hyperactivity, indicating an absence of stimulant
activity (See, FIG. 31). Further evidence for a lack of
psychostimulant activity in Compound B is given in Example 13.
Example 13
Lack of Psychostimulant-Like Effect
[0206] Drug discrimination is an operant paradigm that enables
assessment of drug abuse liability (Yasar & Bergman, 1994.
Amphetamine-like effect of 1-deprenyl (selegiline) in drug
discrimination studies. Clin. Pharmacol. Therap. 56 (S78),
768-773). In this paradigm, psychostimulant properties of compounds
may be determined in rats trained to discriminate methamphetamine
from saline. Hungry rats are initially placed in a test apparatus
where they learn that pressing either of two levers results in
delivery of a food pellet. Once lever pressing has been
established, rats learn that if they are pretreated with
methamphetamine, they must now choose (for example) the left hand
lever in order to obtain food. On other days, rats are pretreated
with vehicle, and must then select the opposite lever in order to
obtain food. In this way, rats must learn to use the interoceptive
cues generated by the psychostimulant drug to guide its choice of
levers. Once lever pressing to a predetermined criterion has been
established, a test compound can be administered. On these days,
pressing either lever results in food, permitting examination of
whether the rats select the methamphetamine or the saline lever. If
the rat chooses the methamphetamine lever, the test drug is said to
have shown stimulus generalization; that is, it is perceived to be
methamphetamine-like by the rat. After administration of Compound B
(0.1-3.2 mg/kg i.p. or 0.32-10 mg/kg p.o.), rats did not select the
methamphetamine lever, indicating a lack of stimulus
generalization. These findings suggest that Compound B may possess
wake promoting activity without psychostimulant drug abuse
liability.
EQUIVALENTS
[0207] While this invention has been particularly shown and
described with references to preferred embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
scope of the invention encompassed by the appended claims.
* * * * *